title: Construction and expression of recombinant fusion protein of thioredoxin-ApoO.
abstract: OBJECTIVE
  To construct human apolipoprotein O (apolipoprotein O, ApoO) expression vector and obtain recombinant fusion protein thioredoxin (Trx)-ApoO by pET prokaryotic expression system.
  
  
  METHODS
  The ApoO gene fragment from the human liver cDNA library was amplified by PCR. The resulting product was cloned into pET-32a(+) vector and sequenced. The confirmed cDNA was cloned into plasmid E.coli DH10B and then transformed into E.coli BL 21 (DE3) where it was induced to express protein by isopropyl β-D-1-thiogalactopyranoside (IPTG). The fusion protein was purified by Ni-NTA resin.
  
  
  RESULTS
  The ApoO gene was cloned by PCR and a 519 bp DNA fragment was shown on the agarose electrophoresis. The cloned gene was sequenced and demonstrated to have the same sequence as that of human ApoO gene in GenBank which justified a successful construction of recombinant plasmid. ApoO cDNA gene fragment was induced by IPTG, and a 34 kD recombinant fusion protein Trx-ApoO was tested on sodium dodecyl sulfate polyacrylamide (SDS-PAGE).
  
  
  CONCLUSIONS
  Human ApoO gene is successfully cloned and its recombinant fusion protein Trx-ApoO is expressed.
authors.affiliation: Department of Cardiovasology, Second Xiangya Hospital, Central South University, Changsha 410011, China.
authors.forename: Chenlu
authors.initials: C
authors.lastname: Wu
authors.forename: Shuiping
authors.initials: S
authors.lastname: Zhao
authors.forename: Bilian
authors.initials: B
authors.lastname: Yu
authors.forename: Dan
authors.initials: D
authors.lastname: Xiong
mesh_terms: D001053:Apolipoproteins / Q000096:biosynthesis* / Q000235:genetics; D003001:Cloning, Molecular; D004926:Escherichia coli / Q000235:genetics / Q000378:metabolism; D005822:Genetic Vectors / Q000235:genetics*; D006801:Humans; D011993:Recombinant Fusion Proteins / Q000096:biosynthesis* / Q000235:genetics; D013879:Thioredoxins / Q000096:biosynthesis* / Q000235:genetics
chemical_list: D001053:Apolipoproteins; D011993:Recombinant Fusion Proteins; D013879:Thioredoxins